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Team:University College London/LabBook/Week3
From 2012.igem.org
Monday (18.6.12)
Aim – Day 1 of Generating Competent Cells: In order to culture our BioBricks in a cell line, we must first enable our cells to uptake plasmid. Today we will be starting the first day of the protocol, whereby we streak the cells on a minimal agar.
In order to culture our BioBricks in a cell line, we must first enable our cells to uptake plasmid. Today we will be starting the first day of the protocol, whereby we streak the cells on a minimal agar.
Method
Step 1 – Setting up Plates: Pour several plates with minimal agar.
Step 2 – Streaking: Streak the chosen cell line onto the minimal agar plate.
Step 3 – Incubation: Incubate at 37oC overnight
Tuesday (19.6.12)
Aim – Picking Colonies: Today we check the colony formation from yesterday’s culture, and pick a colony for inoculation
Method
Step 1 -Creating culture media: In a sterile environment, set up a falcons, with 5mls of Lysogeny Broth and 100ul 1M MgS04.
Step 2 – Selecting a Colony: Select a clear, isolated colony and using an inoculation hoop, scoop up a colony onto the tip. Deposit in the falcon tube
Step 3 - Cell culture: Culture your falcon tubes overnight at a temperature of 37 oC. Leave for no longer than 16 hours.
Wednesday (20.6.12)
Aim – Permeabilisation: Incubating Cells in the presences of CaCl2 will prepare the cell wall, such that it becomes permeable to DNA.
Method
(LOGO) – Day 3 of Generating Competent Cells
